A novel rhodopsin-based voltage indicator for simultaneous two-photon optical recording with GCaMP in vivo.

Autor:	Villette V; Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France., Yang S; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA., Valenti R; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA., Macklin JJ; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA., Bradley J; Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France., Mathieu B; Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France., Lombardini A; Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France., Podgorski K; Allen Institute for Neural Dynamics, Seattle, WA 98109, USA., Dieudonné S; Institut de Biologie de l'École Normale Supérieure (IBENS), École Normale Supérieure, CNRS, INSERM, PSL Research University, Paris 75005, France., Schreiter ER; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA., Abdelfattah AS; Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA 20147, USA.; Department of Neuroscience, Brown University, Providence, RI 02906, USA.; Carney Institute for Brain Science, Brown University, Providence, RI 02906, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Nov 17. Date of Electronic Publication: 2024 Nov 17.
DOI:	10.1101/2024.11.15.623698
Abstrakt:	Genetically encoded voltage indicators (GEVIs) allow optical recording of membrane potential from targeted cells in vivo . However, red GEVIs that are compatible with two-photon microscopy and that can be multiplexed in vivo with green reporters like GCaMP, are currently lacking. To address this gap, we explored diverse rhodopsin proteins as GEVIs and engineered a novel GEVI, 2Photron, based on a rhodopsin from the green algae Klebsormidium nitens . 2Photron, combined with two photon ultrafast local volume excitation (ULoVE), enabled multiplexed readout of spiking and subthreshold voltage simultaneously with GCaMP calcium signals in visual cortical neurons of awake, behaving mice. These recordings revealed the cell-specific relationship of spiking and subthreshold voltage dynamics with GCaMP responses, highlighting the challenges of extracting underlying spike trains from calcium imaging. Competing Interests: Competing interests R.V, E.R.S. and A.S.A have filed patent applications on Voltage Indicators for Two-Photon Imaging
Databáze:	MEDLINE
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